We conducted a systematic study on the reactivity of[Ru_(2)Cp_(2)(CO)4](Cp=η^(5)-C_(5)H_(5))with isocyanides and the subsequent methylation reaction to produce[Ru_(2)Cp_(2)(CO)2(μ-CO){μ-CNMe(R)}]^(+)complexes as CF...We conducted a systematic study on the reactivity of[Ru_(2)Cp_(2)(CO)4](Cp=η^(5)-C_(5)H_(5))with isocyanides and the subsequent methylation reaction to produce[Ru_(2)Cp_(2)(CO)2(μ-CO){μ-CNMe(R)}]^(+)complexes as CF_(3)SO_(3)^(−)salts,[2a-h]^(+)[R=Me,cyclohexyl(Cy),2,6-C_(6)H_(3)Me_(2)(Xyl),1H-indol-5-yl,2-naphthyl,4-C_(6)H_(4)OMe,(S)-CHMe(Ph),CH_(2)Ph(Bn)].The resulting products,including five novel ones,underwent structural characterization by IR and multinuclear NMR spectroscopy,with five of them further confirmed via single crystal X-ray diffraction.Compounds[2a-e,h]CF_(3)SO_(3)exhibit appreciable water solubility,substantial amphiphilic character and out-standing stability in physiological-like solutions(negligible degradation after 72 hours in DMEM at 37℃).Representative complexes[2b]^(+)and[2c]^(+)were additionally characterized through cyclic voltammetry in CH_(2)Cl_(2)and in aqueous phosphate buffer solution.Compounds[2a-d]CF_(3)SO_(3)were assessed for in vitro cyto-toxicity against A2780,A2080R and MCF-7 human cancer cell lines,and[2a-c]CF_(3)SO_(3)revealed significant-to-moderate cytotoxicity,outperforming cisplatin in several cases.The most favourable IC_(50)values were observed for[2d]CF_(3)SO_(3),ranging from 3.7 to 13.0μM.Experiments on the noncancerous human cell line MRC-5 highlighted a reasonable selectivity for[2b-d]CF_(3)SO_(3),with the highest selectivity indexes(SI)calcu-lated as 10.1(ratio of IC_(50)on MRC-5/IC_(50)on A2780)and 8.5(ratio of IC_(50)on MRC-5/IC_(50)on A2780R)for[2d]CF_(3)SO_(3).Subsequently,[2d]CF_(3)SO_(3)was tested across a panel of HOS,A549,PANC1,CaCo2,PC3 and HeLa cancer cells,showing variable cytotoxicity with IC_(50)values in the range of 9.7 to 20.3μM.The cellular effects of[2d]^(+)on A2780 cells were investigated using flow cytometry assays,focusing on the cell cycle modification,time-resolved cellular uptake,intracellular ROS production,mitochondrial membrane depolarization,induction of cell death through apoptosis,activation of caspases 3/7 and induction of autophagy.Overall,the results suggest a diphasic mechanism of action for[2d]^(+),inducing metabolic stress and arresting proliferation in the first/fast phase,followed by the induction of apoptosis and autophagy in the second/slower phase.展开更多
In this work,in situ luminescence analysis was applied for the first time for monitoring the phase transitions of calcium phosphate(CaP)and confirmed by synchrotron in situ X-ray diffraction in addition to in situ inf...In this work,in situ luminescence analysis was applied for the first time for monitoring the phase transitions of calcium phosphate(CaP)and confirmed by synchrotron in situ X-ray diffraction in addition to in situ infrared spectroscopy,with simultaneous measurements of pH and ion conductivity.Applying doped Ce^(3+)and Eu^(3+)as local coordination sensors,the high sensitivity of their emission spectra upon the changes in the coordination sphere of the doped cation sites enabled to detect the formation of amorphous calcium phosphate(ACP)and Ca_(5)(PO_(4))_(3)OH,besides their subsequent transitions to CaHPO_(4)·2H_(2)O and Ca_(8)H_(2)(PO_(4))_(6)·5H_(2)O under real reaction conditions.Calcium phosphates are widely present in mammals and understanding their phase transitions is important to comprehend the conversion between healthy and diseased tissues.In situ luminescence measurements are advantageous for allowing monitoring these phase transitions in a fast and sensitive fashion also in conventional laboratories,independent of synchrotron radiation.展开更多
基金L.B.,T.F.and F.M.thank the University of Pisa(Fondi di Ateneo 2020 and PRA_2020_39)for financial supportJ.V.,T.M.and Z.T.acknowledge the financial support from the ERDF/ESF Project Nanotechnologies for Future(CZ.02.1.01/0.0/0.0/16_019/0000754)thank Ms Marta Rešováfor help with biological testing,Prof.MarekŠebela for assistance with MALDI-TOF MS experiments,and Dr Ondřej Vrobel for assist-ance with some of the mass spectrometry experiments。
文摘We conducted a systematic study on the reactivity of[Ru_(2)Cp_(2)(CO)4](Cp=η^(5)-C_(5)H_(5))with isocyanides and the subsequent methylation reaction to produce[Ru_(2)Cp_(2)(CO)2(μ-CO){μ-CNMe(R)}]^(+)complexes as CF_(3)SO_(3)^(−)salts,[2a-h]^(+)[R=Me,cyclohexyl(Cy),2,6-C_(6)H_(3)Me_(2)(Xyl),1H-indol-5-yl,2-naphthyl,4-C_(6)H_(4)OMe,(S)-CHMe(Ph),CH_(2)Ph(Bn)].The resulting products,including five novel ones,underwent structural characterization by IR and multinuclear NMR spectroscopy,with five of them further confirmed via single crystal X-ray diffraction.Compounds[2a-e,h]CF_(3)SO_(3)exhibit appreciable water solubility,substantial amphiphilic character and out-standing stability in physiological-like solutions(negligible degradation after 72 hours in DMEM at 37℃).Representative complexes[2b]^(+)and[2c]^(+)were additionally characterized through cyclic voltammetry in CH_(2)Cl_(2)and in aqueous phosphate buffer solution.Compounds[2a-d]CF_(3)SO_(3)were assessed for in vitro cyto-toxicity against A2780,A2080R and MCF-7 human cancer cell lines,and[2a-c]CF_(3)SO_(3)revealed significant-to-moderate cytotoxicity,outperforming cisplatin in several cases.The most favourable IC_(50)values were observed for[2d]CF_(3)SO_(3),ranging from 3.7 to 13.0μM.Experiments on the noncancerous human cell line MRC-5 highlighted a reasonable selectivity for[2b-d]CF_(3)SO_(3),with the highest selectivity indexes(SI)calcu-lated as 10.1(ratio of IC_(50)on MRC-5/IC_(50)on A2780)and 8.5(ratio of IC_(50)on MRC-5/IC_(50)on A2780R)for[2d]CF_(3)SO_(3).Subsequently,[2d]CF_(3)SO_(3)was tested across a panel of HOS,A549,PANC1,CaCo2,PC3 and HeLa cancer cells,showing variable cytotoxicity with IC_(50)values in the range of 9.7 to 20.3μM.The cellular effects of[2d]^(+)on A2780 cells were investigated using flow cytometry assays,focusing on the cell cycle modification,time-resolved cellular uptake,intracellular ROS production,mitochondrial membrane depolarization,induction of cell death through apoptosis,activation of caspases 3/7 and induction of autophagy.Overall,the results suggest a diphasic mechanism of action for[2d]^(+),inducing metabolic stress and arresting proliferation in the first/fast phase,followed by the induction of apoptosis and autophagy in the second/slower phase.
基金the development of the in situ analysis software as well as the DFG(Priority Program 1415,project BE 1653/29-1,project TE 1147/1-1),Daimler and Benz Foundation and MATsynCELL for the financial support.
文摘In this work,in situ luminescence analysis was applied for the first time for monitoring the phase transitions of calcium phosphate(CaP)and confirmed by synchrotron in situ X-ray diffraction in addition to in situ infrared spectroscopy,with simultaneous measurements of pH and ion conductivity.Applying doped Ce^(3+)and Eu^(3+)as local coordination sensors,the high sensitivity of their emission spectra upon the changes in the coordination sphere of the doped cation sites enabled to detect the formation of amorphous calcium phosphate(ACP)and Ca_(5)(PO_(4))_(3)OH,besides their subsequent transitions to CaHPO_(4)·2H_(2)O and Ca_(8)H_(2)(PO_(4))_(6)·5H_(2)O under real reaction conditions.Calcium phosphates are widely present in mammals and understanding their phase transitions is important to comprehend the conversion between healthy and diseased tissues.In situ luminescence measurements are advantageous for allowing monitoring these phase transitions in a fast and sensitive fashion also in conventional laboratories,independent of synchrotron radiation.
